High glucose inhibits human epidermal keratinocyte proliferation for cellular studies on diabetes mellitus.
High glucose inhibits human epidermal keratinocyte proliferation for cellular studies on diabetes mellitus.
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DOI：
10.1111/j.1742-4801.2005.00148.x
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发表时间：
2005-12-01
影响因子：
3.1
通讯作者：
Marcelo, Cynthia L
中科院分区：
文献类型：
--
作者：
Terashi, Hiroto;Izumi, Kenji;Marcelo, Cynthia L
为了更清楚地阐明糖尿病中延迟的伤口愈合，我们在6mM（对照组）和12mM葡萄糖（高糖组）的“完全”MCDB 153培养基中培养人表皮角质形成细胞。高脂血症减慢了它们的增殖速度，抑制了它们的DNA合成和总蛋白的产生。高糖组原代培养1个月后细胞停止增殖，而对照组细胞生长超过40天。高糖组平均群体倍增时间为5.43 d（对照组为3.65 d，P = 0.02），高糖组平均群体倍增时间为5.27 d（对照组为7.25 d，P = 0.001）。他们指出，长期暴露于高葡萄糖降低体外人表皮角质形成细胞的复制寿命。此外，用薄层色谱和气相色谱法分析膜磷脂中的脂肪酸含量表明，在两种条件下培养的角质形成细胞之间没有差异。葡萄糖转运蛋白1的免疫细胞化学染色显示，高糖组28.1%的细胞表达葡萄糖转运蛋白1，几乎是对照组13.2%的2倍（P = 0.008）。高葡萄糖对表皮角质形成细胞的不良影响的机制目前尚不清楚，但它表明高血糖症对葡萄糖代谢的任何直接影响的可能性，而不改变细胞膜上的脂质代谢。本研究所建立的高血糖组可作为研究糖尿病皮肤表皮病变的体外模型。
In order to more clarify the delayed wound healing in diabetes mellitus, we cultured the human epidermal keratinocytes in both 6 mM (control group) and 12 mM glucose (high-glucose group) of "complete" MCDB 153 medium. Hyperglycaemia slowed the rate of their proliferation and inhibited their DNA synthesis and the production of total proteins. By 1 month after primary seeding in high-glucose group, the cells ceased their proliferation, whereas the cells in control group grew for more than 40 days. Mean population doublings in high-glucose group was 5.27 (vs. 7.25 in control, P = 0.001), and mean population doubling time during 1 month in high glucose group was 5.43 days (vs. 3.65 days in control, P = 0.02). They indicate that prolonged exposure to high glucose decreases the replicative life span of human epidermal keratinocytes in vitro. Furthermore, analysis of fatty acid contents in membrane phospholipids with thin-layer and gas chromatography showed no difference between the cultured keratinocytes in both conditions. Immunocytochemical staining of glucose transporter 1 shows that 28.1% of cells in high-glucose group were almost twice positive of those in control group (13.2%, P = 0.008). The mechanism of the ill effects of high glucose on epidermal keratinocytes is not so far clear, but it indicates the possibility of any direct effect of hyperglycaemia on glucose metabolism without changing lipid metabolism on cell membrane. The high-glucose group presented in this report can be available as an in vitro valuable study model of skin epidermal condition on diabetes mellitus.